Applications of magnetic beads with covalently attached oligonucleotides in hybridization: isolation and detection of specific measles virus mRNA from a crude cell lysate.
A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to magnetic beads (Dynabeads) has been developed. The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 M guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA.
Albretsen
C
C
Center of Biotechnology, University of Bergen, Norway.
Kalland
K H
KH
Haukanes
B I
BI
Håvarstein
L S
LS
Kleppe
K
K
D001483:Base Sequence; D008280:Magnetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids / Q000302:isolation & purification / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification
D009696:Nucleic Acids; D009841:Oligonucleotides; D012333:RNA, Messenger; D012367:RNA, Viral